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Figure 3 | Journal of Hematology & Oncology

Figure 3

From: Soluble NKG2D ligand promotes MDSC expansion and skews macrophage to the alternatively activated phenotype

Figure 3

sMIC promotes induction of MDSC through engagement of NKG2D and activation of STAT3. (a) Bone marrow cells from B6 mice were co-cultured with sMICB expressing prostate tumor cell line TRAMP-C2 (TC2-sMICB-GFP, also as TC2-sMICB) or control TC2-GFP cells at a ratio of 10:1 for 3 days. Cells were harvested and stained with antibodies specific to CD45 (to differentiate myeloid cells from tumor cells), Gr-1, and CD11b for flow cytometry analyses. Data show representative plots and summary of CD11b+Gr-1+ in gated CD45+ cells. (b) Bone marrow cells from B6 mice were cultured in the presence of GM-CSF, with various concentration of purified sMICB for 3 days. Cells were harvested and analyzed for CD11b and Gr-1 expression. Data show representative plots and summary of the percentage of cells that are CD11b+Gr-1+. (c) Representative histogram showing NKG2D expression on myeloid cells with various culture conditions. Grey-filled profile, staining with isotype control. Open profile, staining with anti-NKG2D antibody CX5. (d) Bone marrow cells from B6 mice were cultured with GM-CSF in the presence or absence of sMICB (25 ng/ml) and with and without the NKG2D blocking antibody CX5 or STAT3 inhibitor AG490. Data show representative plots and summary of CD11b+Gr-1+ cells after 3 days of culture. (e) Levels of intracellular phosphorylated STAT3 in bone marrow myeloid cells cultured in the condition as described in (d). Three replicates were performed in each experiment. Data represent five independent experiments. *P < 0.05.

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